immunoblotting

Summary

Summary: Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.

..Overall, our findings show that although APOBEC family members are highly related, they exhibit significantly distinct antiviral characteristics that may provide new insights into host-pathogen interactions...

The physical relationship between infectivity and prion protein aggregates is strain-dependent

..In particular, nuclear proteins and proteins involved in regulating metabolic processes have high phosphorylation site occupancy in mitosis. This suggests that these proteins may be inactivated by phosphorylation in mitotic cells...

Akt forms an intracellular complex with heat shock protein 90 (Hsp90) and Cdc37 and is destabilized by inhibitors of Hsp90 function

Andrea D BassoProgram in Pharmacology, Weill Graduate School of Medical Sciences, Cornell University and the Program in Cell Biology and Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10021, USAJ Biol Chem 277:39858-66. 2002

..Thus, transduction of growth factor signaling via the Akt and Raf pathways requires functional Hsp90 and can be coordinately blocked by its inhibition...

Anti-inflammatory compounds parthenolide and Bay 11-7082 are direct inhibitors of the inflammasome

..Our results thus elucidate the molecular mechanism for the therapeutic anti-inflammatory activity of parthenolide and identify vinyl sulfones as a new class of potential therapeutics that target the NLRP3 inflammasome...

James HarrisDepartment of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, University of New Mexico School of Medicine, 915 Camino de Salud NE, Albuquerque, NM 87131, USAImmunity 27:505-17. 2007

..These findings establish a mechanism through which Th1-Th2 polarization differentially affects the immune control of intracellular pathogens...

Sir2-dependent activation of acetyl-CoA synthetase by deacetylation of active lysine

..We conclude that neither bortezomib nor metformin has significant benefit in this native Tsc2(+/-) mouse model, which suggests limited benefit of these compounds in the treatment of TSC hamartomas and related lesions...

..diagnostic sensitivities of the three analytical techniques (enzyme-linked immunosorbent assay [ELISA], immunoblotting, and PCR) were compared by using a group of patients (n = 19) with suspected ocular toxoplasmosis...

Integrin-specific activation of Rac controls progression through the G(1) phase of the cell cycle

..This process is mediated by SOS and PI-3K and requires coordinate upstream signals through Shc and FAK. These results indicate that Rac is a crucial mediator of the integrin-specific control of cell cycle in endothelial cells...

Use of a Dictyostelium model for isolation of genetic loci associated with phagocytosis and virulence in Klebsiella pneumoniae

..Therefore, we have identified genes involved in resistance to phagocytosis in K. pneumoniae using Dictyostelium, and this model is useful to explore genes associated with resistance to phagocytosis in heavily encapsulated bacteria...

..used to characterize the reactivity of EBA sera and to map the epitopes on the murine antigen by ELISA and immunoblotting. The patients' autoantibody binding to murine collagen VII triggered pathogenic events as demonstrated by a ..

..In contrast, expression of a catalytically inactive hSir2 protein potentiates p53-dependent apoptosis and radiosensitivity. We propose that hSir2 is involved in the regulation of p53 function via deacetylation...

Validity and reproducibility of measurement of islet autoreactivity by T-cell assays in subjects with early type 1 diabetes

..Hence we postulate that STT3B-dependent posttranslational N-glycosylation is part of a triage-salvage system recognizing cryptic N-glycosylation sites of secretory proteins to preserve protein homeostasis...

Yi He LingDepartment of Oncology, Albert Einstein College of Medicine, Bronx, New York 10461, USA New York University Cancer Institute, School of Medicine, New York University, New York, New York 10016, USAJ Biol Chem 278:33714-23. 2003

..Thus ROS generation plays a critical role in the initiation of the bortezomib-induced apoptotic cascade by mediation of the disruption of Delta psi m and the release of cytochrome c from mitochondria...

Liver-specific deletion of the NADPH-cytochrome P450 reductase gene: impact on plasma cholesterol homeostasis and the function and regulation of microsomal cytochrome P450 and heme oxygenase

Jun GuWadsworth Center, New York State Department of Health, and School of Public Health, State University of New York, Albany, New York 12201, USAJ Biol Chem 278:25895-901. 2003

..The Alb-Cre/Cprlox mouse represents a unique model for studying the in vivo function of hepatic HO and microsomal CYP-dependent pathways in the biotransformation of endogenous and xenobiotic compounds...

..Furthermore, the link we observed between siRNA efficacy and phenotypic outcome indicates that distinct stages during cell cycle progression are disrupted by the differential modulation of KIF14 expression...

..The effect of EGF to regulate ZPR1 binding is dependent on tyrosine phosphorylation of the EGFR. ZPR1 therefore represents a prototype for a class of molecule that binds to the EGFR and is released from the receptor after activation...

O-linked β-N-acetylglucosamine transferase is indispensable in the failing heart

..These data provide keen insights into the pathophysiology of the failing heart and illuminate a previously unrecognized point of integration between metabolism and cardiac function in the failing heart...

..Thus, our data unravel the molecular organization of the ubiquitination complex that catalyzes p27 ubiquitination and provide unique insights into the specificity of substrate recognition by SCF(Skp2)...

..Specific antisera directed towards the carboxyl terminal of the rac protein kinase were prepared and used to identify that phosphorylated several substrates in immunoprecipitates prepared with the rac-specific antisera...

..This approach can be used to study other signaling systems and suggests that the p53-Mdm2 feedback loop generates a 'digital' clock that releases well-timed quanta of p53 until damage is repaired or the cell dies...

Activation of innate immune antiviral responses by Nod2

Ahmed SabbahDepartment of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, USANat Immunol 10:1073-80. 2009

..Thus, the function of Nod2 as a viral PRR highlights the important function of Nod2 in host antiviral defense mechanisms...

..In addition, this study implicates ATR as a direct upstream activator of Chk1 in human cells...

An IcmF family protein, ImpLM, is an integral inner membrane protein interacting with ImpKL, and its walker a motif is required for type VI secretion system-mediated Hcp secretion in Agrobacterium tumefaciens

..Together, these results suggest that the formation of the inner rod, not the needle, is critical for substrate specificity switching and that YscP and YscU exert their effects on substrate export by controlling the secretion of YscI...

..These studies provide compelling evidence for a prominent role of Cav-1 in regulating the functional activity of not only 5-HT(2A) serotonin receptors but also selected Galpha(q)-coupled receptors...

..The delayed transactivation of the C/EBPalpha and PPARgamma genes by C/EBPbeta appears necessary to allow mitotic clonal expansion, which would otherwise be prevented, because C/EBPalpha and PPARgamma are antimitotic...

..of BMP-2 in activating these important transcription factors leading to osteoclast differentiation using immunoblotting, immunohistochemistry, in situ hybridization, ChIP analysis and qRTPCR techniques...

..Concentrations and phosphorylation status of selected proteins will be determined by immunoblotting. Immunohistochemistry will be used to assess muscle fiber hypertrophy, fiber types, numbers of fibers, and ..

..migration and development of PH with a variety of cell culture, transfection, siRNA, immunoprecipitation and immunoblotting techniques and with the use of normal and transgenic rodent model experiments...

..are attached to streptavidin-coated beads to capture the PICs from extracts and analyze their composition by immunoblotting. The functions of the PIC are studied using histone modification/remodeling assays and in vitro transcription...

..NFS-sId and control NFS-N mice will be utilized for all experiments. Gene expression and immunoblotting assays, combined with in vivo immunoprecipitation experiments, will shed light on the influence of quantity, ..

..cells will be incubated in trophoblast CM and secretion and phosphorylation of IGFBP-1 (Western blot, 2-D immunoblotting, ELISA and mass spectrometry approaches) as well as IGF-I secretion (ELISA) will be determined...

..1b. The mechanism of cholinergic agonist-induced cell proliferation will be studied using immunoblotting to probe for activated proteins in the p44/42 MARK, p38 MARK and JNK pathways, and by examining the roles of ..

..The purified fractions will be analyzed by 2-dimensional gel electrophoresis and immunoblotting. The internal amino acid sequences will be obtained after proteolytic digestion of th purified protein...

..known to have anti-70k antibodies from our large well-characterized cohort of patients using oxidative 70k and immunoblotting. Specificity will be determined in blocking studies using molar excess of either intact, apoptotic 70k or ..

..receptor content in subcellular fractions isolated by differential centrifugation will be quantitated using immunoblotting with specific anti-LDL receptor antibodies, LDL receptors in subcellular fractions will be quantitated using ..